title: β2 adrenergic receptor fluorescent protein fusions traffic to the plasma membrane and retain functionality.
abstract: Green fluorescent protein (GFP) has proven useful for the study of protein interactions and dynamics for the last twenty years. A variety of new fluorescent proteins have been developed that expand the use of available excitation spectra. We have undertaken an analysis of seven of the most useful fluorescent proteins (XFPs), Cerulean (and mCerulean3), Teal, GFP, Venus, mCherry and TagRFP657, as fusions to the archetypal G-protein coupled receptor, the β2 adrenergic receptor (β2AR). We have characterized these β2AR::XFP fusions in respect to membrane trafficking and G-protein activation. We noticed that in the mouse neural cell line, OP 6, that membrane bound β2AR::XFP fusions robustly localized in the filopodia identical to gap::XFP fusions. All β2AR::XFP fusions show responses indistinguishable from each other and the non-fused form after isoprenaline exposure. Our results provide a platform by which G-protein coupled receptors can be dissected for their functionality.
authors.affiliation: Department of Biological Sciences, Hunter College and The Graduate Center Biochemistry, Biology and Biopsychology and Behavioral Neuroscience Programs, CUNY, New York, New York, United States of America.
authors.forename: Jaclyn
authors.initials: J
authors.lastname: Bubnell
authors.forename: Patrick
authors.initials: P
authors.lastname: Pfister
authors.forename: Maria L
authors.initials: ML
authors.lastname: Sapar
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Rogers
authors.forename: Paul
authors.initials: P
authors.lastname: Feinstein
mesh_terms: D000818:Animals; D019390:Arrestins / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D004705:Endocytosis / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007545:Isoproterenol / Q000494:pharmacology; D008024:Ligands; D051379:Mice; D021381:Protein Transport / Q000187:drug effects; D011554:Pseudopodia / Q000187:drug effects / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013997:Time Factors; D014168:Transferrin / Q000378:metabolism; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D008024:Ligands; D018343:Receptors, Adrenergic, beta-2; D011993:Recombinant Fusion Proteins; D014168:Transferrin; D000071557:beta-Arrestins; D049452:Green Fluorescent Proteins; D007545:Isoproterenol